GSM7837932: Normal, scRNAseq; Homo sapiens; RNA-Seq
A single-cell suspension of kidney cells was prepared using Multi Tissue Dissociation Kit 2 (#130-110-203, Miltenyi Biotec) according to the manufacturer's instructions. Kidneys were harvested from anesthetized mice, cut into quarters, and transferred into the gentleMACS C Tube containing the enzyme mix. After digesting the tissues using gentleMACS Dissociator, they were passed through a 70-μm and 40-μm cell strainer, sequentially, and washed with ice-cold PBS to obtain the single-cell suspension. Red blood cells were lysed using 10×RBC lysis buffer (#00-4300, eBioscience).Single cells isolated from the IRI kidneys at indicated time points were stained with Zombie NIR viability dye (BioLegend); CD45- and CD 45+ live cells were sorted using BD FACS AriaTM cell sorter. FACS-sorted cells were resuspended to a ﬁnal cell concentration of 700–1200 cells/μL with > 80% viability as determined by LUNA-FL™ Automated Fluorescence Cell Counter (Logos Biosystems). RNA libraries  were prepared with Chromium Single cell 3' Reagent v3 Kits according to the manufacturer's protocol.